Methods of using humanized antibodies and compositions for binding sphingosine-1-phosphate

Abstract

The present invention relates to anti-S1P agents, particularly humanized monoclonal antibodies (and antigen binding fragments thereof) specifically reactive with S1P, compositions containing such antibodies (or fragments), and the use of such antibodies (or fragments), for example, to treat diseases and conditions associated with aberrant levels of S1P.

1. A method of treating a disease or disorder correlated with an aberrant level of S1P, comprising administering to a subject known or suspected to be suffering from a disease or disorder correlated with an aberrant level of sphingosine-1-phosphate (S1P) a therapeutically effective amount of an isolated humanized antibody, or an antigen binding fragment thereof, that binds S1P to effect treatment of the disease or disorder, wherein the antibody or an antigen binding fragment thereof comprises at least one heavy chain variable domain and at least one light chain variable domain, wherein:
 A. each heavy chain variable domain comprises the following:
 (i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO: 13); 
 (ii) a second sequence of amino acid residues selected from the group consisting of AISPRHDITKYNEMFRG (SEQ ID NO: 31) and CISPRHDITKYNEMFRG (SEQ ID NO:14), provided that if the second sequence of amino acid residues is CISPRHDITKYNEMFRG (SEQ ID NO:14), the heavy chain variable domain is humanized; and 
 (iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO: 15); and 
 
 B. each light chain variable domain comprises the following:
 (i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO: 10); 
 (ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO: 11); and 
 (iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO: 12). 
 
 
     
     
       2. A method according to  claim 1 , wherein the disease or disorder is a cancer. 
     
     
       3. A method according to  claim 1 , wherein the disease or disorder is a cerebrovascular disease. 
     
     
       4. A method according to  claim 1 , wherein the disease or disorder is a cardiovascular disease. 
     
     
       5. A method according to  claim 1 , wherein:
 A. each heavy chain variable domain of the antibody or an antigen binding fragment thereof comprises a heavy chain variable domain comprising a sequence of amino acid residues having an amino acid sequence EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLEWMG AISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMYFCARG GFYGSTIWFDFWGQGTMVTVSS (SEQ ID NO: 32, residues 20-140, inclusive); and 
 B. each light chain variable domain of the antibody or an antigen binding fragment thereof comprises a light chain variable domain comprising a sequence of amino acid residues having an amino acid sequence ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPGKAPKLLISEG NILRPGVPSRFSSSGYGTDFTLTISKLQPEDFATYYCLQSDNLPFTFGQGTK LEIK (SEQ ID NO: 33, residues 21-127, inclusive). 
 
     
     
       6. A method according to  claim 1 , wherein:
 A. each heavy chain of the antibody or an antigen binding fragment thereof comprises a sequence of amino acid residues having an amino acid sequence: EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLEWMG AISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMYFCARG GFYGSTIWFDFWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPG (SEQ ID NO: 38, residues 20-454, inclusive); and 
 B. each light chain of the antibody or an antigen binding fragment thereof comprises a sequence of amino acid residues having an amino acid sequence: ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPGKAPKLLISEG NILRPGVPSRFSSSGYGTDFTLTISKLQPEDFATYYCLQSDNLPFTFGQGTK LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC (SEQ ID NO: 37, residues 21-234, inclusive). 
 
     
     
       7. A method according to  claim 1 , wherein:
 A. each heavy chain variable domain of the antibody or an antigen binding fragment thereof comprises the same heavy chain variable domain amino acid sequence as the heavy chain variable domain encoded by the heavy chain gene in vector pATH1009 in ATCC Accession No. PTA-8421; and 
 B. each light chain variable domain of the antibody or an antigen binding fragment thereof comprises the same light chain variable domain amino acid sequence as the light chain variable domain encoded by the light chain gene in vector pATH1009 in ATCC Accession No. PTA-8421. 
 
     
     
       8. A method according to  claim 1 , wherein:
 A. each heavy chain of the antibody or an antigen binding fragment thereof comprises the same heavy chain amino acid sequence as the heavy chain of the antibody expressed by CHO cell line LH1 275, as deposited under accession number ATCC PTA-8422; and 
 B. each light chain of the antibody or an antigen binding fragment thereof comprises the same light chain amino acid sequence as the light chain of the antibody expressed by CHO cell line LH1 275, as deposited under accession number ATCC PTA-8422. 
 
     
     
       9. A method according to  claim 1 , wherein at least one amino acid residue of the antibody or antigen binding fragment thereof is glycosylated. 
     
     
       10. A method according to  claim 1 , wherein the composition comprises an isolated humanized antibody that includes two heavy chains and two light chains. 
     
     
       11. A method of treating a disease or disorder correlated with an aberrant level of S1P, comprising administering to a subject known or suspected to be suffering from a disease or disorder correlated with an aberrant level of S1P a therapeutically effective amount of an isolated humanized antibody, or an antigen binding fragment thereof, specifically reactive with S1P to effect treatment of the disease or disorder, wherein the antibody or an antigen binding fragment thereof comprises at least one heavy chain variable domain and at least one light chain variable domain, wherein:
 A. each heavy chain variable domain comprises:
 (i) a sequence of amino acid residues having an amino acid sequence EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLE WMGAISPRHDITKYNEMFRGQVTISADKSSSTAYLQWSSLKASDT AMYFCARGGFYGSTIWFDFWGQGTMVTVSS (SEQ ID NO: 32, residues 20-140, inclusive); or 
 (ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of A(i), above provided such sequence of amino acid residues comprises a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO: 13), a second sequence of amino acid residues selected from the group consisting of AISPRHDITKYNEMFRG (SEQ ID NO: 31) and CISPRHDITKYNEMFRG (SEQ ID NO:14), and a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO: 15); and 
 
 B. each light chain variable domain comprises:
 (i) a sequence of amino acid residues having an amino acid sequence ETTVTQSPSFLSASVGDRVTITCITTTDIDDDMNWFQQEPG KAPKLLISEGNILRPGVPSRFSSSGYGTDFTLTISKLQPEDF ATYYCLQSDNLPFTFGQGTKLEIK (SEQ ID NO: 33, residues 21-127, inclusive); or 
 (ii) a sequence of amino acid residues having at least about 80% sequence identity to the amino acid sequence of B(i), above provided such sequence of amino acid residues comprises a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO: 10), a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO: 11), and a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO: 12). 
 
 
     
     
       12. A method according to  claim 11 , wherein the disease or disorder is a cancer. 
     
     
       13. A method according to  claim 11 , wherein the disease or disorder is a cerebrovascular disease. 
     
     
       14. A method according to  claim 11 , wherein the disease or disorder is a cardiovascular disease. 
     
     
       15. A method according to  claim 1 , wherein:
 A. each heavy chain variable domain of the antibody or an antigen binding fragment thereof comprises:
 (i) a first sequence of amino acid residues of sequence DHTIH (SEQ ID NO: 13); 
 (ii) a second sequence of amino acid residues AISPRHDITKYNEMFRG (SEQ ID NO: 31); and 
 (iii) a third sequence of amino acid residues of sequence GGFYGSTIWFDF (SEQ ID NO: 15); and 
 
 B. each light chain variable domain of the antibody or an antigen binding fragment thereof comprises:
 (i) a first sequence of amino acid residues of sequence ITTTDIDDDMN (SEQ ID NO: 10); 
 (ii) a second sequence of amino acid residues of sequence EGNILRP (SEQ ID NO: 11); and 
 (iii) a third sequence of amino acid residues of sequence LQSDNLPFT (SEQ ID NO: 12). 
 
 
     
     
       16. A method according to  claim 1 , wherein each heavy chain of the antibody or an antigen binding fragment thereof comprises a sequence of amino acid residues having an amino acid sequence: EVQLVQSGAEVKKPGESLKISCQSFGYIFIDHTIHWMRQMPGQGLEWMGAISPRHDITK YNEMFRGQVTISADKSSSTAYLQWSSLKASDTAMYFCARGGFYGSTIWFDFWGQGTM VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 38, residues 20-454, inclusive). 
     
     
       17. A method according to  claim 11 , wherein at least one amino acid residue of the antibody or antigen binding fragment thereof is glycosylated. 
     
     
       18. A method according to  claim 11 , wherein the composition comprises an isolated humanized antibody that includes two heavy chains and two light chains.